Preparation And Characterization Of The Sulfated Fenugreek Gum And Preliminary Study Of Inhibition Of Tumor Cell Activity In Vitro

Fenugreek?Trigonella foenum-graecum L.? is a leguminous herb. Its seeds can be used as medicine in the seed endosperm rich in polysaccharides, also known as fenugreek gum or fenugreek polysaccharide?fenugreek gum, FG?, as the main component of galactomannan?Galactomannan?, with blood sugar regulation,anti-tumor, antioxidant etc. biological activity, widely used in pharmaceutical,chemical, health products and food industry. In recent years, through the introduction of various functional groups?such as sulfate, phosphate etc.? to enhance plant polysaccharides activity has become a hot research topic in the field of natural products, including sulfated polysaccharide derivatives because of their excellent physical and chemical properties the biological activity and received special attention.Studies have shown that the sulfate radical polysaccharide chain of different substituted positions?C-6, C-2 or C-3? to the biological activity of different synthesis,with different degree of substitution and substitution Set of activity of polysaccharide derivatives has major significance. Therefore, the paper is based on the material of FG,through non- and Regioselectivities for two synthetic methods of preparing the different structural characteristics of the gourd Pakistan polysaccharide sulfate and RSFG SFG, and their in vitro antitumor activity was studied. The main conclusions are as follows:1.The response surface method to optimize the preparation process and structure characterization of the polysaccharide sulfate?SFG?By chlorosulfonic acid pyridine method?CSA/Py? of fenugreek polysaccharide sulfate?SFG? was synthesized by the response surface?RSM? experiments to determine the optimum reaction conditions were: the ratio of CSA/Py to 1:2.3, the reaction time is 4.3 h, the reaction temperature is 64 ?, the degree of substitution?DSS? between 0.42⁰.56, conforming to the model through expectations. Infrared spectroscopy?FT-IR?, X ray photoelectron spectroscopy?XPS?, the product was characterized with 13 C NMR, sulfuric acid groups have been introduced in the SFe structure, S exists in the form of S6+, mainly to replace the position in C-6. Size exclusion chromatography coupled with light scattering system?SEC-LLS? to determine the average molecular weight?MW? showed that acidic reaction conditions cause the reduction of MW, molecular weight distribution narrows,the higher the DSS, the smaller the MW and methylene blue?MB? by sulfate complexation experiments confirmed that polysaccharide solution conformation changes, atomic force microscope?AFM? was also observed in linear branched molecular morphology modification reaction After the short chain, indicating a greater degree of structure sulfation of polysaccharides.2. The regional selective synthesis and structural characterization of the polysaccharide sulfate?RSFe?In acidic conditions polysaccharide sulfation restrictions in the choice of sterically hindered silane agent has better stability to acid?two tert butyl phenyl chlorosilane, TBDPSCl? selective protection of polysaccharide primary hydroxyl groups were regioselective sulfation of Fe. For the unimolecular nucleophilic substitution?SN1? Rules activation, solvent effect and polysaccharide primary hydroxyl group of the key factors affecting the reaction, the reaction conditions were optimized and analyzed the possible reaction mechanism; according to the characteristics of polysaccharide silicon ether intermediates, design and screening of different sulfated reaction parameters and selective removal of protection system and condition, fenugreek polysaccharide sulfate synthesis area selective?RSFe?, and the product of the reaction steps were characterized. The determination of DSS was 0.29,the total yield was 111.4%.FT-IR, XPS, 13 C CP/MAS NMR analysis showed that the sulfate group in RSFe place mainly in C-2 or C-3 position, and the product obtained by the non selective synthesis process is obviously different.3.A preliminary study on the antitumor activity of SFe and Fe in vitroAfter the sulfated derivatives were synthesized successfully, and Fe was studied with different DS, sulfated derivatives of MW?SFe₇, SFe₁2 and SFe₁3? on human lung cancer cell?A549?, human cervical cancer cells?Hela? and human hepatocellular carcinoma cells?Hep G-2? inhibited the in vitro effect on cell cycle and regulation dead. Sulfation of SFe₇, Hela and Hep G-2 of tumor cells IC50 values were 171.9 and196.1 ?g/m L, SFe₁₃ on Hela and Hep G-2 tumor cells IC50 values were 181.14 and 200?g/m L, compared to all kinds of plant polysaccharides sulfuric acid ester and reported in the literature, showing good anti tumor in vitro the role of.Fe showed inhibition of tumor cells in vitro, stronger on A549, Hela and Hep G-2 of tumor cells IC50 values were 85.04120.01 and 148.77 ?g/m L, but in the cell cycle and apoptosis in the experiment, Fe was not found in a cycle resistance of A549 obviously and Hep G-2tumor cells The effects of stagnation and promoting apoptosis should be studied in the future, and the mechanism of action of different antitumor pathways is elucidated.